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Neural stem cell injection for treating brain damage disease

A technology of neural stem cells and brain injury, applied in the direction of nervous system cells, animal cells, nervous system diseases, etc., can solve the problems of no specific treatment for children with cerebral palsy, can not achieve treatment, etc., achieve good stemness and differentiation potential, in vitro The effect of strong amplification ability

Inactive Publication Date: 2017-05-10
SHANGHAI ANGECON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, most of the drugs for the treatment of children with cerebral palsy are to provide nutrition and relieve symptoms, and cannot achieve the purpose of treatment.
In other words, there is currently no effective drug for the specific treatment of children with cerebral palsy on the market.

Method used

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  • Neural stem cell injection for treating brain damage disease
  • Neural stem cell injection for treating brain damage disease
  • Neural stem cell injection for treating brain damage disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Preparation of primary neural stem cells

[0053] 1. Induction of primary neural stem cells from embryonic stem cells (ESCs)

[0054] Use a cell scraper to pick ESCs cell clones, blow them gently, and then resuspend them with neural stem cell induction medium, transfer them to an ultra-low adsorption culture dish for suspension culture for about 3 days, collect the formed cell balls, blow them gently, and then use neural stem cell The stem cell induction medium was resuspended, suspended and cultured in an ultra-low adsorption culture dish for 3 days, and the formed cell spheres were collected; the culture dish coated with Laminin was washed twice with DPBS without Ca and Mg one day in advance, and the collected The cell balls were resuspended with the complete medium of neural stem cells, inoculated in the coated culture dish for adherent culture, and the medium was changed every 2-3 days until obvious neural-like colonies were formed, and then the formed ce...

Embodiment 2

[0068] Example 2: Preparation of P1-P5 generation neural stem cells

[0069] 1. Passage and culture of primary neural stem cells

[0070] Inject the prepared 10ml sterile neural digestion solution into the centrifuge tube containing the primary neural stem cell spheres with a sterile syringe. Seal the mouth of the centrifuge tube with parafilm and let stand for 3 minutes. Add the prepared sterilized nerve tissue digestion solution, put it into a carbon dioxide incubator at 35°C for 30 minutes (manually shake for 5 seconds every 10 minutes). Low-temperature centrifuge at 4°C, 300g, and centrifuge for 10 minutes. Discard the supernatant with a sterile disposable pipette, add 1-5mL DMEM / F12 to rinse with a high-power pipette and pipette, and blow up and down 2-3 times to separate the tissue. Low-temperature centrifuge at 4°C, 300g, and centrifuge for 10-20 minutes. Add 1-5mL hibernation medium again for washing, and wash three times in total. Discard the supernatant with a s...

Embodiment 3

[0091] Example 3 Preparation of P6-P9 generation neural stem cells

[0092] 1. Recovery of cells

[0093] Take out the frozen cells from the liquid nitrogen tank, put them on the resuscitation rack in the electric heating constant temperature water tank (the water temperature is kept at about 40 ℃), shake quickly to rewarm. Resuscitation is complete when the solid changes to a liquid.

[0094] 1.1 Cell washing

[0095] Seal the mouth of the centrifuge tube with parafilm and let stand for 3 minutes. Add the prepared sterilized nerve tissue digestion solution, put it into a carbon dioxide incubator at 35°C for 5-60 minutes (manually shake for 5 seconds every 10 minutes). Low-temperature centrifuge at 4°C, 300g, and centrifuge for 10 minutes.

[0096] 1.2 Sampling counting and washing

[0097] Discard the supernatant with a sterile disposable pipette, add 1-10mL DMEM / F12 to rinse with a high-power pipette and pipette, and blow up and down 2-3 times to separate the tissue. L...

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Abstract

The invention relates to a neural stem cell injection, which is derived from a human fetal cerebral cortex source and is used for treating various brain damages. The neural stem cell injection at least comprises 1*10<6> neural stem cells, wherein the neural stem cells are obtained by separately culturing waste fetal cerebral cortex tissues; the neural stem cell are cultured by tissue separation and primary cell culture, purification and amplification culture of the neural stem cells, building of a neural stem cell library and preparation of the neural stem cell injection. The cells can be amplified by 5000 times in a serum-free culture system. According to the use condition of the cells, P5-generation neural stem cells of seed cells are recovered and counted, and undergo suspension culture in a serum-free culture medium and digestion passage, till P9-generation neural stem cells are obtain by repeating the previous steps. The neural stem cell injection is 95 percent in purity, and can be applied to treatment of various brain damage diseases such as cerebral apoplexy, hypoxic-ischemic brain damage and infantile cerebral palsy.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a neural stem cell injection for treating brain injury diseases, a preparation method and a use method thereof. Background technique [0002] Cerebral palsy in children usually refers to central movement disorders caused by non-progressive brain damage or abnormal brain development caused by various reasons before birth to one month after birth. Clinically, it is characterized by abnormal posture and muscle tone, muscle weakness, involuntary movement, and ataxia, often accompanied by sensory, cognitive, communication, behavioral and other disorders and secondary skeletal muscle abnormalities, and may have seizures. There is no ideal treatment for this disease at present, the disability rate is high, and the mental burden and economic burden caused to the patient himself and his family are heavy. [0003] According to the data released by the Ministry of Health, there are 16 million ne...

Claims

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Application Information

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IPC IPC(8): A61K35/30A61K9/08A61P25/00A61P9/10C12N5/0797
CPCA61K9/0019A61K9/0043A61K9/0085A61K9/08A61K35/30C12N5/0623C12N2500/30C12N2500/32C12N2501/11C12N2501/115C12N2501/235
Inventor 郑佳威赵雄飞黄倩莹
Owner SHANGHAI ANGECON BIOTECH
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